中国氏族医药杂志 2024年8月第25卷第8期37实验研究HPLC法测定大鼠血浆中CYP450不同亚型探针药物浓度“张东I耶国栋\郝海梅2甄海涛2常乐2张艳东2曹晓峰2(1.包头医学院基础医学与法医学院，内蒙古包头014060;2.包头医学院药学院，内蒙古包头014060)摘 要：目的：建立同时测定细胞色素P450 (cytochrome P450.CYP450)亚型CYP1A2、CYP2C9和CYP2E1活性的 Cocktail探针药物溶液的高效液相色谱(HPLC)检测方法。方法：分别选择茶碱、甲苯磺丁腺和氯哇沙宗作为CYP1A2,
CYP2C9和CYP2E1的特异性探针药物，根据其理化特点配制成Cocktail组合探针溶液。使用HPLC梯度洗脱法同时测定3
种探针药物的血药浓度，并对其方法学进行验证。结果：本研究建立的HPLC法可以同时测定大鼠血浆中3种探针药物的
浓度，各探针药物鸽吸收峰分离完全，无杂峰干扰;线性关系良好，线性范围为0.02~50|xg/mL,最低检测限为0.02?g/mL。
日内、日间精密度均小于10%,回收率在87.74%~112.39%之间，稳定性高，室温放置、-20咒冷冻1周和3周的RSD均小于
10%?将该方法应用于研究大鼠体内此3种酶的活性，证明其具有较好的灵敏度。结论：经方法学验证，此法可同时测定
3种探针药物茶碱、甲苯磺丁腺和氯哇沙宗的血浆浓度。关键词：CYP450; Cocktail探针药物法；HPLC中图分类号:R969.1 文献标识码:A 文章编号:1006-6810(2024 )08-0037-04Determination of probe concentration of CYP450 isoforms in plasma of rats by HPLCZHANG Dong1, WU Guo-dong2* , HAO Hai-mei2, ZHEN Hai-tao2, CHANG Le2, ZHANG Yan-dong2, CAO Xiao-feng2(1. School of basic and forensic medicine,Baotao Medical College,Bao Tou 014060,China;2. School of Pharmacy,Baotou Medical College,Bao Tou 014060,China)[Abstract ] Objective: To establish an HPLC method for simultaneously determination of the metabolic activities of cytochrome P450 (CYP450) isoforms CYP1A2, CYP2C9和CYP2E1. Methods: Cocktail probe solution was prepared by mixing
3 specific probe drugs of CYP1A2, CYP2C9 and CYP2E1, namely theophylline, tolbutamide and chlorzoxazone respectively.An
HPLC method for simultaneous determination of the above 3 probes was established and validated. Results: An HPLC method for simultaneous determination of the plasma concentrations of 3 probe drugs was established.The absorption peak of each probe was separated without interference of impurity peak.For each probe, the linear range was 0.02~50|Xg/mL, the minimum detection limit was 0.02pig/mL, the intraday and interday precisions were less than 10%, the recovery was between87.74%-
112.39%, the RSD of stability after frozen 1 and 3 weeks was less than 10%. The method showed good sensitivity for pharmacokinetic study in vivo. Conclusion: The established HPLC method is efficient and reliable for simultaneous determination of theophylline, tolbutamide and chlorzoxazone, thus can be applied to activity evaluation of CYP1A2, CYP2C9
and CYP2E1 with a cocktail approach.[Key words ] CYPP450; Cocktail Probe Drugs; HPLC目前，临床上许多疾病的治疗均采用联合用药，这样就 代谢动力学层面。药物代谢动力学层面上的相互作用可发
很可能导致药物在体内发生相互作用，就会导致不良反应
生在药物在体内过程的任一环节，比如吸收、分布、代谢或
的发生。药物相互作用可分为药物效应动力学层面和药物 者是排泄过程。其中发生在药物在体内代谢过程的相互作
用占药物代谢动力学层面相互作用的40%,发生率很高，临
△基金项目：包头医学院科学研究基金项目扬帆计划(No.BYJJ- 床意义也非常重大叫细胞色素P450(CYP450)是生物体内
YF 201618)；内蒙古自治区自然科学基金面上(联合)项目(No. 参与各类内源性和外源性物质代谢的主要酶系，是由许多
2017MS(LH )0808)作者简介:张东，副教授,硕士。研究方向：中蒙药药理学。E-mail：
同工酶组成的超家族叫这些同工酶可以代谢人体内90%以
zdong696@ 163.com。上的药物。CYP1A2、2C和2E1在整个CYP450大家族中占
*通讯作者:郭国栋,副教授，硕士。研究方向:天然药物提取分离O 的比例比较高(CYP2E1大概占7%,CYP1A2大概占13%,
E-mail: wgdzd@ 126.com。CYP2C大概占20%)叫CYP450酶的活性容易受外源性物
HPLC法测定大鼠血浆中CYP450不同亚型探针药物浓度 - 论文
